Development of novel gene therapy based on the analysis of anti-apoptotic signals in ovarian cancer

基于卵巢癌抗凋亡信号分析的新型基因疗法的开发

基本信息

批准号：
11470347
负责人：
KONISHI Ikuo
金额：
$ 9.47万
依托单位：
SHINSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B).
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11470347/
关键词：
ovarian cancer apoptosis gene therapy adenovirus gonadotropin insulin-like growth factor-1 p53 Bax

项目摘要

Among the gynecological malignant tumors, ovarian carcinoma is most frequently associated with chemoresistance and poor prognosis of the patients. The effect of chemotherapy with various anti-cancer agents including cisplatin is shown to be mediated by intracellular apoptotic signals. Anti-apoptotic factors, either extrinsic or intrinsic, may influence the chemotherapeutic effect, and may result in the chemoresistance of ovarian cancer cells. Therefore, the objective of our study is to analyze the anti-apoptotic signals in ovarian carcinoma cells, and to develop novel gene therapy targeting these anti-apoptotic signals.Our study demonstrated that an approximately half of ovarian carcinomas expressed LH/hCG receptor at both mRNA and protein levels. In ovarian cancer cell line OVCAR3 expressing LH/hCG receptors, hCG inhibited cisplatin-induced apoptosis via up-regulation of IGF-1. IGF-1 also inhibited cisplatin-induced apoptosis and a neutralizing antibody to IGF-1 receptor restored the … More apoptosis. Wortmannin, an inhibitor of phosphatidyl inositol 3-kinase, blocked the anti-apoptotic effect of IGF-1. Therefore, gonadotropin is an important extrinsic factor of anti-apoptotic signal in ovarian cancer cell. Treatment with recombinant adenovirus expressing anti-sense IGF-1 mRNA was effective for restoring the cisplatin-induced apoptosis.Abnormality of p53 tumor suppressor gene is present in more than half of ovarian carcinomas. Pro-apoptotic bax gene is under the regulation of p53, and p53 abnormality resulted in chemoresistance as an intrinsic anti-apototic factor. In vitro, Bax protein expression was attenuated in cisplatin-resistant cell lines such as A2780/cDDP (p53 mutated) and SKOV3 (p53 deleted), compared with cisplatin-sensitive A2780 (p53 wild type). We developed recombinant adenovirus which highly expresses the Bax protein. The Bax protein was successfully induced by treatment with the Bax/adenovirus transfer, and apoptotic effect was obtained in the cisplatin-resistant ovarian cancer cells. Therefore, adenovirus-mediated introduction of Bax may be a useful gene thrapy in the treatment of chemoresistant ovarian carcinomas. Less

在妇科恶性肿瘤中，卵巢癌最常与患者的化学耐药性和预后不良有关。对包括顺铂（顺铂）的各种抗癌药物的化学疗法的作用被证明是由细胞内凋亡信号介导的。抗凋亡因子（外在的或内在的）可能会影响化学治疗作用，并可能导致卵巢癌细胞的化学耐药性。因此，我们研究的目的是分析卵巢癌细胞中的抗凋亡信号，并开发针对这些抗凋亡信号的新型基因疗法。在表达LH/HCG受体的卵巢癌细胞系OVCAR3中，HCG通过上调IGF-1抑制了顺铂诱导的凋亡。 IGF-1还抑制了顺铂诱导的细胞凋亡，对IGF-1受体的中和抗体恢复了……更多的凋亡。 Wortmannin是一种磷脂酰肌醇3-激酶的抑制剂，阻断了IGF-1的抗凋亡作用。因此，促性腺激素是卵巢癌细胞中抗凋亡信号的重要外部因素。用表达抗敏感性IGF-1 mRNA的重组腺病毒治疗可有效恢复顺铂诱导的凋亡。p53肿瘤抑制基因的含量在超过一半以上的卵巢癌中呈现。促凋亡的BAX基因受p53的调节，p53异常导致化学抗性作为固有的抗脑病因子。在体外，与顺铂敏感的A2780（p53野生型）相比，在顺铂耐药细胞系（p53突变）和SKOV3（p53删除）中，Bax蛋白表达被减弱。我们开发了重组腺病毒，该腺病毒高度表达了Bax蛋白。通过用BAX/腺病毒转移处理成功诱导了Bax蛋白，并在抗顺铂的卵巢癌细胞中获得了凋亡作用。因此，腺病毒介导的BAX的引入可能是化学耐药性卵巢癌的有用基因攻击。较少的